| ObjectiveThis study is aim to reveal the correlation between autophagy and pathogenesis of rheumatoid arthritis by analysing expression of PI3K/akt/m TOR and Beclin1,ATG5,ATG 8 from peripheral blood mononuclear cells in patients with rheumatoid arthritis.Methods1.The patients were selected from The First Affiliated Zhangzhou Hospital of Fujian Medical University in January 2014 to June 2015.60 patients were diagnosed as rheumatoid arthritis based on RA classification diagnostic criteria from the American rheumatism association in 2009.The RA disease activity index(DAS28)were divided into three groups: low activity(DAS28 < 3.2,group L,22cases),moderate activity(3.2< DAS2<8 5.1,group M,20cases)and high activity(DAS28 > 5.1,group H,18cases).25 cases were selected as healthy control(N group).Peripheral blood were collected.2.Detection of serological index: ESR was detected by fμLly automatic blood sedimentation instrument,RF and hs-CRP were detected by fμLl automatic immune turbidimetric instrument and anti-CCP antibody was detected by ELISA.3.Gene expression level of Beclin1,ATG5,ATG8 from autophagy related genes and PI3 K,Akt,m TOR from PI3K/Akt/m TOR signaling pathways were detected with FQ-PCR and western blot respectively.Results1.Each group RA patients serum ESR,hs-CRP,RF,anti CCP results:1.1 low(L),in the activities of groups group(M),high activity(H)serum ESR value were 9.67 ±6.22mm/h,57.38 ± 26.62mm/h,50.80 ± 26.80mm/h were significantly higher than that of healthy controls(3.90 ±1.21 mm/h),P values were 0.023,0.001,0.002,less than 0.05,statistically significant.In group(M)(H),high activity group was obviously higher than that of group(L),P values were 0.009,0.010;the group(M)with(H),P value is 0.056.There was no significant difference.1.2 each index of the hs-CRP in serum in patients with RA:L、M、H group serum hs-CRP value were 1.12 ±1.21 IU/ml,26.73±16.30 IU/ml,31.16 ± 9.77 IU/ml,which were higher than healthy controls 0.63±0.23.IU/ml,P values were 0.039,0.001,0.002,less than 0.05,which had statistically significant.In group(M)(H),was obviously higher than that of groups(L).1.3 each RF index in serum in patients with RA: group(L)、(M)、(H)serum RF value of groups were383.72±167.80mg/L,142.33±126.02mg/L,233.75±170.96,mg/L which were higher than healthy controls 2.81±1.76mg/L,P values were 0.001,0.020,0.009,less than 0.05,which had statistically significant.1.4 each CCP resistance index in serum in patients with RA:group(L)、(M)、(H)serum anti CCP values were 963.13±617.85RU/ml,468.79±329.39RU/ml,202.45 ±119.33RU/ml,which were higher than healthy controls 5.13±2.47RU/ml,P values were 0.001,0.013,0.015,less than 0.05,which had statistically significant.group(L)in the above group(M),group(H),and will decrease.1.5 ESR,hs-CRP,RF,anti CCP four joint diagnostic effect: RA sensitivity to RF was 87%,specificity was 79%.The sensitivity of ESR,hs-CRP,anti CCP and RF combined detection of RA was 68%,the specificity was 100%.2.Groups of RA patients with PBMC of AKT,PI3 K,m TOR,ATG5,ATG8,Beclin1 m RNA expression level:2.1 The m RNA of PI3 K in group(L),y group(M),group(H)values were 2.04 ±0.74,4.05± 0.46,2.18 ±0.25,which were higher than healthy controls 0.49±0.13,P values were 0.020,0.009,0.019,less than 0.05,statistically significant.Activities in the group(M)is higher thanthe group(H)(L)of high activity,P were 0.018,0.031.2.2 The m RNA of AKT in group(L),medium activity group(M)and group(H)were 0.46±0.32,0.83±0.42,0.65±0.38,which were higher than the healthy control 0.23 + 0.21.P values were 0.030,0.017,0.027,less than 0.05,had statistically significant.2.3 The m RNA of m TOR in group(L),group(M)and group(H)were 0.94 ± 0.12,1.12 ± 0.23 and 0.23 ±0.30,which were higher than healthy controls 0.32 ± 0.06.P values were 0.003,0.002,0.001,less than 0.05,had statistically significant2.4 each PBMC ATG5 m RNA expression level in patients with RA: The m RNA of ATG5 ingroup(L),group(M)3.38±0.45,4.46±1.28,which were higher than healthy controls 2.06±0.25,P were 0.035,0.013,less than 0.05,statistically significant.The group(H)was 2.36 ± 0.96 Which is no difference with control group(p = 0.078).2.5 groups of PBMC ATG8 m RNA expression level in patients with RA: The m RNA of ATG8 in group(L),group(M)and group(H)were 1.98±0.28,1.58±0.81,were higher than healthy controls 0.49±0.09,P were 0.001,0.002,0.001,less than 0.05,statistically significant.Activities in the group(M)was below the group(L)and group(H),P value were 0.044,0.043.2.6 groups of PBMC Beclin1 m RNA expression level in patients with RA: The m RNA of Beclin1 in group(L),(M),(H)were 2.08±0.66,2.19±0.30 which were higher than healthy controls 0.76±0.10,P values were 0.012,0.013,0.015,less than 0.05,had statistically significant.2.7 P13 K and AKT,m TOR,Beclin1,bivariate regression were correlation between ATG5、ATG8、P13K and AKT.P13 K and ATG8 which had negativelycorrelated,and with the m TOR,Beclin1,ATG5 were positively correlated.3.Each group P13 K,AKT,m TOR,ATG8 protein in expression :3.1 The expression of PI3 K protein of PBMC in RA patients: compared with healthy controls,the group(L),group(M)andgroup(H)RA in peripheral blood mononuclear cells of patients with lower expression of PI3 K,were0.44±0.11、0.14±0.02、0.06±0.01(P=0.008,0.005,0.001),the difference was statistically significant.3.2The expression of AKT of PBMC in RA patients: compared with healthy controls,the group(L),group(M)and group(H)RA in peripheral blood mononuclear cells of patients with the increase of AKT protein expression were 2.51±0.11、18.29±4.52、15.13±3.28(P=0.035,0.001,0.002).The difference was statistically significant.3.3The expression of m TOR protein of PBMC in RA patients : compared with the healthy control,group(M),group(H)RA in peripheral blood mononuclear cells of patients with the increase of m TOR protein expression were 1.5±7.52、35.13±8.62(P=0.002,0.001),the difference was statistically significant.The low activity group(L)1.25±0.13 and the control group had no significant difference.3.4The expression of ATG8 protein of PBMC in RA patients : compared with healthy controls,thegroup(L),group(M)and group(H)RA in peripheral blood mononuclear cells of patients with the increase of ATG8 protein expression were 1.67±0.23、2.92±0.82、11.08±2.32(P=0.027,0.020,0.001),the difference was statistically significant.Conclusions1.ESR,hs-CRP,RF,anti CCP can help diagnosision of RA patients and have reference value to assess the severity,activity and prognosis of the disease.2.The signaling pathway of PI3K/akt/m TOR signaling pathway which participate in the autophagy of rheumatoid arthritis,which may be related to the severity of RA.3.The expression changes of Beclin1 and Atg8 in peripheral blood mononuclear cells of RA patients have a certain correlation with the prognosis and activity of RA. |
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